Stable isotope labelling with amino acids in cell culture for human embryonic stem cell proteomic analysis

Linda Harkness; Tatyana A Prokhorova; Moustapha Kassem; Blagoy Blagoev

doi:10.1007/978-1-61779-794-1_20

Stable isotope labelling with amino acids in cell culture for human embryonic stem cell proteomic analysis

Linda Harkness, Tatyana A Prokhorova, Moustapha Kassem, Blagoy Blagoev

Research output: Contribution to journal › Journal article › Research

Abstract

The identification and quantitative measurements of proteins in human embryonic stem cells (hESC) is a fast growing interdisciplinary area with an enormous impact on understanding the biology of hESC and the mechanism controlling self-renewal and differentiation. Using a quantitative mass spectroscopic method of stable isotope labelling with amino acids during cell culture (SILAC), we are able to analyse differential expression of proteins from different cellular compartments and to identify intracellular signalling pathways involved in self-renewal and differentiation. In this chapter, we provide a detailed method for creating SILAC media suitable for use in hESC experiments, additionally we describe methods for the isolation of membrane fractions and cytosolic and nuclear/membrane fractions.

Original language	English
Journal	Methods in Molecular Biology
Volume	873
Pages (from-to)	297-305
Number of pages	9
ISSN	1064-3745
DOIs	https://doi.org/10.1007/978-1-61779-794-1_20
Publication status	Published - 2012

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title = "Stable isotope labelling with amino acids in cell culture for human embryonic stem cell proteomic analysis",

abstract = "The identification and quantitative measurements of proteins in human embryonic stem cells (hESC) is a fast growing interdisciplinary area with an enormous impact on understanding the biology of hESC and the mechanism controlling self-renewal and differentiation. Using a quantitative mass spectroscopic method of stable isotope labelling with amino acids during cell culture (SILAC), we are able to analyse differential expression of proteins from different cellular compartments and to identify intracellular signalling pathways involved in self-renewal and differentiation. In this chapter, we provide a detailed method for creating SILAC media suitable for use in hESC experiments, additionally we describe methods for the isolation of membrane fractions and cytosolic and nuclear/membrane fractions.",

author = "Linda Harkness and Prokhorova, {Tatyana A} and Moustapha Kassem and Blagoy Blagoev",

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AU - Harkness, Linda

AU - Prokhorova, Tatyana A

AU - Kassem, Moustapha

AU - Blagoev, Blagoy

PY - 2012

Y1 - 2012

N2 - The identification and quantitative measurements of proteins in human embryonic stem cells (hESC) is a fast growing interdisciplinary area with an enormous impact on understanding the biology of hESC and the mechanism controlling self-renewal and differentiation. Using a quantitative mass spectroscopic method of stable isotope labelling with amino acids during cell culture (SILAC), we are able to analyse differential expression of proteins from different cellular compartments and to identify intracellular signalling pathways involved in self-renewal and differentiation. In this chapter, we provide a detailed method for creating SILAC media suitable for use in hESC experiments, additionally we describe methods for the isolation of membrane fractions and cytosolic and nuclear/membrane fractions.

AB - The identification and quantitative measurements of proteins in human embryonic stem cells (hESC) is a fast growing interdisciplinary area with an enormous impact on understanding the biology of hESC and the mechanism controlling self-renewal and differentiation. Using a quantitative mass spectroscopic method of stable isotope labelling with amino acids during cell culture (SILAC), we are able to analyse differential expression of proteins from different cellular compartments and to identify intracellular signalling pathways involved in self-renewal and differentiation. In this chapter, we provide a detailed method for creating SILAC media suitable for use in hESC experiments, additionally we describe methods for the isolation of membrane fractions and cytosolic and nuclear/membrane fractions.

U2 - 10.1007/978-1-61779-794-1_20

DO - 10.1007/978-1-61779-794-1_20

M3 - Journal article

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JO - Methods in Molecular Biology

JF - Methods in Molecular Biology

SN - 1064-3745

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